To determine the affinity for d-serine or glutamate in GluN2B- and GluN2B(P553T)-transfected cells, concentration-response curves were constructed from whole-cell currents elicited by rapid jumps of 0.5-s duration at different concentrations of the coagonist d-serine (10−9 to 10−4 M) in the presence of 1 mM glutamate or at different concentrations of the agonist glutamate (10−8 to 10−2 M) in the presence of 1 mM d-serine. Concentration-response curves were fitted individually for every cell using the Hill equationI=Imax1+(EC50[A])nHwhere Imax is the maximum current, [A] is the concentration of d-serine, nH is the slope (Hill) coefficient, and EC50 is the concentration of d-serine or glutamate that produces a half-maximum response. Each data point was then normalized to the maximum response obtained in the fit. The average of the normalized values with their SEM were plotted together and fitted again with the Hill equation. The minimum and maximum values were constrained to asymptote 0 and 1, respectively.

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